Search results for "Fluorescent"

showing 10 items of 863 documents

Effects of drought and re-watering on water transport in Sedum sediforme (Jacq.) Pau

2011

Settore BIO/03 - Botanica Ambientale E ApplicataSettore BIO/04 - Fisiologia Vegetalewater transport Sedum drought stress fluorescent tracers hydraulic conductance tylosis
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Functional studies of regulatory genes in the sea urchin embryo.

2009

Settore BIO/11 - Biologia Molecolaremicroinjection sea urchin embryo gene function cis-regulatory analysis green fluorescent protein
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Metilazione del DNA in artrite reumatoide

2005

Lo stato di metilazione del DNA genomico e del gene PTHrP è stato valutato con tecniche molecolari e citogenetiche in artrite reumatoide (AR), patologia autoimmune caratterizzata anche da alta incidenza di linfomi e da ipercalcemia per overespressione del gene PTHrP. La metilazione del DNA, infatti, ha un ruolo critico nello sviluppo delle malattie neoplastiche; il gene PTHrP avendo tre promotori uno dei quali contiene un’isola CpG è un buon candidato per la deregolazione da alterato pattern di metilazione locale. Le indagini sulla metilazione genomica, condotte su DNA estratto da sangue periferico di pazienti e di donatori e amplificato in reazioni di Methylation-Sensitive Arbitrarily Prim…

Settore BIO/18 - Geneticainstead chromosomes of controls were almost uniformly decorated by brilliant grains. Studies on methylation of PTHrP gene promoter 2 performed on five CpG island internal sites using the Methylation-Sensitive Restriction Endonuclease Multiplex (MSREM)-PCR showed that one of the sites nearest the trascription starting point is heavy methylated in a significantly high number of RA patients. Thus RA seems to be characterized by genomewide hypomethylation associated with local hypermethylation like the most part of tumors. This result raises the possibility that susceptibility to lymphomas is related to abnormal DNA methylation levels and suggests the opportunity to evaluate the DNA methylation status in RA patientin fact the demethylating therapies together with diet and life style can act towards an increase of tumor risk. Future studies using a larger number of subjects could confirm these findings.Rheumatoid Arthritis (RA) is a chronic multisystem inflammatory disease characterized by high recurrence of lymphomas as well as hypercalcemia due to PTHrP overexpression. Because of DNA methylation plays a critical role in development of neoplasias we determined in RA patients the global DNA methylation status and local methylation pattern of the CpG island of one of the three promoters of PTHrP gene utilizing molecular and cytogenetic techniques. Investigations performed on DNA from peripheral blood of patients and donors amplified by Methylation-Sensitive Arbitrarily Primed (MeS-AP)-PCR indicated that RA is strongly associated with global DNA hypomethylation. Similarly chromosomal DNA methylation pattern analysis by indirect immunofluorescence technique with anti 5-methylcitosine antibody showed all peripheral lymphocyte metaphases from RA patients with chromosomes weakly fluorescent without discrete grain
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Suitability of two rapid lateral flow immunochromatographic assays for predicting SARS‐CoV‐2 neutralizing activity of sera

2020

Purpose: Assessment of commercial SARS-CoV-2 immunoassays for their capacity to provide reliable information on sera neutralizing activity is an emerging need. We evaluated the performance of two commercially-available lateral flow immunochromatographic assays (LFIC) (Wondfo SARS-CoV-2 Antibody test and the INNOVITA 2019-nCoV Ab test) in comparison with a SARS-CoV-2 neutralization pseudotyped assay for COVID-19 diagnosis in hospitalized patients, and investigate whether the intensity of the test band in LFIC associates with neutralizing antibody (NtAb) titers. Patients and Methods: Ninety sera were included from 51 patients with moderate to severe COVID-19. A green fluorescent protein (GFP)…

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)Green Fluorescent ProteinsAntibodies ViralNeutralizing antibodiesNeutralizationSARS‐CoV‐2Green fluorescent protein03 medical and health sciencesCOVID-19 Testing0302 clinical medicineCOVID‐19VirologyLateral flow 16 immunochromatographic assaysHumansMedicineImmunochromatographic Assays030212 general & internal medicineNeutralizing antibodyResearch ArticlesImmunoassaybiologybusiness.industrySARS-CoV-2COVID-19biology.organism_classificationAntibodies NeutralizingVirologyTiterInfectious DiseasesImmunoglobulin MVesicular stomatitis virusImmunoglobulin GSpike Glycoprotein Coronavirusbiology.proteinlateral flow immunochromatographic assays030211 gastroenterology & hepatologyLteral flow immunochromatographic assaysAntibodybusinessResearch Article
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A novel cell wall protein specific to the mycelial form of Yarrowia lipolytica.

1996

A cDNA clone specifying a cell wall protein was isolated from a Yarrowia lipolytica cDNA library. The cDNA library was constructed in the expression vector lambda gt 11, with the RNA isolated from actively growing mycelial cells. The deduced amino acid sequence shows that the encoded protein contains an N-terminal hydrophobic signal peptide. We have designated this protein YWP1 for Yarrowia lipolytica cell Wall Protein. Northern hybridization identified YWP1 transcript only when Y. lipolytica was growing in the mycelial form. The encoded protein seems to be covalently bound to the glucan cell wall since it is not released from the cell walls by sodium dodecyl sulphate extraction, but it is …

Signal peptideDNA ComplementaryTranscription GeneticHydrolasesBlotting WesternGenetic VectorsMolecular Sequence DataRestriction MappingBioengineeringApplied Microbiology and BiotechnologyBiochemistryCell wallFungal ProteinsOpen Reading FramesTransformation GeneticCell WallComplementary DNAGene Expression Regulation FungalYeastsGeneticsEscherichia coliAmino Acid SequenceCloning MolecularFluorescent Antibody Technique IndirectPeptide sequenceAntibodies FungalGene LibraryExpression vectorbiologyBase SequencecDNA libraryRNASodium Dodecyl SulfateYarrowiaRNA Fungalbiology.organism_classificationBlotting NorthernBlotting SouthernBiochemistrySaccharomycetalesElectrophoresis Polyacrylamide GelBiotechnologyYeast (Chichester, England)
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Altered intracellular sorting signals do not influence the efficacy of genetic melanoma vaccines incorporating helper determinants in mice.

2004

Background A genetic melanoma vaccine consisting of cDNA encoding the model self-antigen tyrosinase-related protein 2 (TRP2) fused in-frame to the immunogenic enhanced green fluorescent protein (EGFP) was able to break immune tolerance and stimulate CD8+ T cells in vivo. In the present study we investigated whether alteration of the intracellular antigen localization as a result of the linkage with immune-enhancing helper proteins affects the resulting immune response. Methods Expression plasmids and recombinant adenoviruses were constructed encoding various fusion proteins with different intracellular sorting signals which direct the antigen to the cytosol, the endoplasmic reticulum or the…

Skin Neoplasmsmedicine.medical_treatmentRecombinant Fusion ProteinsGenetic VectorsGreen Fluorescent ProteinsMelanoma ExperimentalAutoimmunityBiologyCancer VaccinesMelanoma VaccineImmune toleranceMiceImmune systemAntigenDrug DiscoveryGeneticsmedicineAnimalsMolecular BiologyGenetics (clinical)MelanomaELISPOTImmunotherapyGenetic TherapyT-Lymphocytes Helper-Inducermedicine.diseaseMolecular biologyFusion proteinCell biologyIntramolecular OxidoreductasesMice Inbred C57BLProtein TransportCD4 AntigensMolecular MedicineImmunizationThe journal of gene medicine
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Dye selection for live cell imaging of intact siRNA

2011

Abstract Investigations into the fate of small interfering RNA (siRNA) after transfection may unravel new ways to improve RNA interference (RNAi) efficiency. Because intracellular degradation of RNA may prevent reliable observation of fluorescence-labeled siRNA, new tools for fluorescence microscopy are warranted to cover the considerable duration of the RNAi effect. Here, the characterization and application of new fluorescence resonance energy transfer (FRET) dye pairs for sensing the integrity of duplex siRNA is reported, which allows an assessment of the degradation status of an siRNA cell population by live cell imaging. A panel of high-yield fluorescent dyes has been investigated for …

Small interfering RNACell SurvivalClinical BiochemistryPopulationBiologyBiochemistryLive cell imagingRNA interferenceFluorescence Resonance Energy TransferFluorescence microscopeAnimalsRNA Small InterferingeducationMolecular BiologyCells CulturedFluorescent Dyeseducation.field_of_studyMicroscopy ConfocalBrainEndothelial CellsRNATransfectionHydrogen-Ion ConcentrationMolecular biologyRatsFörster resonance energy transferBiophysicsBiological Chemistry
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Efficient selection of silenced primary cells by flow cytometry

2007

Background: RNA interference has emerged as a new and potent tool to knockdown the expression of target genes and to investigate their functions. For short time experiments with mammalian cell lines, RNA interference is typically induced by transfecting small interfering RNAs (siRNAs). Primary cells constitute important experimental systems in many studies because of their similarity to their in vivo counterparts; however, transfection of these cells has been found to be difficult. As a consequence, RNA interference of primary cells may result in mixed phenotypes because of the simultaneous presence in the same preparation of transfected and nontransfected cells. This may be particularly in…

Small interfering RNAHistologyfluorescent siRNAsCell SeparationBiologyTransfectionPathology and Forensic MedicineFlow cytometryRNA interferencemedicineHumansTrypsinGene SilencingRNA Small InterferingGeneCells CulturedGene knockdownMessenger RNAmedicine.diagnostic_testCell BiologyTransfectionCell sortingFlow CytometryMolecular biologyMicroscopy FluorescencemRNA knocking-down in fibroblastsRNAiCytometry Part A
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Flow cytometric kinetic assay of the activity of Na+/H+ antiporter in mammalian cells.

2004

Background The Na+/H+ exchanger (NHE) of mammalian cells is an integral membrane protein that extrudes H+ ion in exchange for extracellular Na+ and plays a crucial role in the regulation of intracellular pH (pHi). Thus, when pHi is lowered, NHE extrudes protons at a rate depending of pHi that can be expressed as pH units/s. Methods To abolish the activity of other cellular pH-restoring systems, cells were incubated in bicarbonate-free Dulbecco's modified Eagle's medium buffered with HEPES. Flow cytometry was used to determine pHi with 2′,7′-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester or 5-(and-6)-carboxy SNARF-1 acetoxymethyl ester acetate, and the appropriate fluo…

Sodium-Hydrogen ExchangersTime FactorsNigericinIntracellular pHBiophysicsIonophoreNaphtholsBiochemistryModels BiologicalPathology and Forensic MedicineFlow cytometryCell Linechemistry.chemical_compoundJurkat CellsMiceEndocrinologyChondrocytesIschemiamedicineExtracellularAnimalsHumansBenzopyransMuscle SkeletalCells CulturedFluorescent DyesHEPESmedicine.diagnostic_testDose-Response Relationship DrugRhodaminesCell BiologyHematologyHydrogen-Ion ConcentrationFlow CytometryFluoresceinsAmilorideKineticsBiochemistrychemistryCell cultureCalibrationNIH 3T3 Cellsmedicine.drugCytometry. Part A : the journal of the International Society for Analytical Cytology
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Aberrant methylation patterns at the two-cell stage as an indicator of early developmental failure

2002

The fertilized mouse egg actively demethylates the paternal genome within a few hours after fertilization, whereas the maternal genome is only passively demethylated by a replication-dependent mechanism after the two-cell stage. This evolutionarily conserved assymetry in the early diploid mammalian embryo may have a role in methylation reprogramming of the two very different sets of sperm and egg chromatin for somatic development and formation of totipotent cells. Immunofluorescence staining with an antibody against 5-methylcytosine (MeC) showed that the incidence of abnormal methylation patterns differs between mouse two-cell embryos from superovulated females, nonsuperovulated matings, an…

Somatic cellCleavage Stage OvumFluorescent Antibody TechniqueSuperovulationAcetaldehydeFertilization in VitroBiologyAndrologyMiceGeneticsmedicineAnimalsBlastocystEnzyme InhibitorsGeneticsEmbryo cultureEmbryoCell BiologyMethylationDNA MethylationSpermmedicine.anatomical_structureembryonic structuresDNA methylationReprogrammingDevelopmental BiologyMolecular Reproduction and Development
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